Purification of a heat-stable activator protein for ADP-ribosylation factor-dependent phospholipase D.
A heat-stable activator for ADP-ribosylation factor (ARF)-dependent phospholipase D (PLD) was purified to near homogeneity from rat kidney cytosol by a sequential column chromatography. The purified activator has a molecular mass of 23 kDa on SDS-PAGE. Using a partially purified ARF-dependent PLD from rat kidney, the activator synergistically stimulates PLD with ARF in time- and dose-dependent manner. In the absence of ARF, the activator has little or no effect. The purified activator also stimulates PLD under several conditions including permeabilized cell system, suggesting that the activator is a physiologically relevant regulator of PLD.
Akisue
T
T
Department of Biochemistry, Kobe University School of Medicine, Japan.
Jinnai
H
H
Hitomi
T
T
Miwa
N
N
Yoshida
K
K
Nakamura
S
S
D020727:ADP-Ribosylation Factors; D000818:Animals; D002463:Cell Membrane Permeability / Q000187:drug effects; D003600:Cytosol / Q000201:enzymology; D004789:Enzyme Activation / Q000502:physiology; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D018922:HL-60 Cells; D006801:Humans; D007668:Kidney / Q000737:chemistry* / Q000201:enzymology; D008970:Molecular Weight; D010739:Phospholipase D / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism; D011506:Proteins / Q000302:isolation & purification* / Q000494:pharmacology*; D051381:Rats; D013301:Streptolysins
D010743:Phospholipids; D011506:Proteins; D013301:Streptolysins; D006160:Guanosine Triphosphate; D010739:Phospholipase D; D019204:GTP-Binding Proteins; D020727:ADP-Ribosylation Factors